研究报告

基于蛋白质组学解析桑椹低聚半乳糖通过碳水化合物代谢调控鼠李糖乳酪杆菌EN-08增殖的机制

  • 陈心如 ,
  • 李倩 ,
  • 庞道睿 ,
  • 邢东旭 ,
  • 邹宇晓 ,
  • 黎尔纳
展开
  • 1(华南农业大学 食品学院,广东 广州,510642)
    2(广东省农业科学院蚕业与农产品加工研究所,农业农村部功能食品重点实验室,广东省农产品加工重点实验室,广东 广州,510610)
第一作者:硕士研究生(黎尔纳副研究员为通信作者,E-mail:lierna@gdaas.cn)

收稿日期: 2025-02-04

  修回日期: 2025-06-01

  网络出版日期: 2026-03-13

基金资助

广东省自然科学基金-青年提升项目(2024A1515030093);国家重点研发计划专项(2023YFF110380504);国家蚕桑产业技术体系建设专项资助(CARS-18-ZJ0503)

Mechanism of mulberry galacto-oligosaccharides regulating proliferation of Lacticaseibacillus rhamnosus EN-08 through carbohydrate metabolism: A proteomics-based analysis

  • CHEN Xinru ,
  • LI Qian ,
  • PANG Daorui ,
  • XING Dongxu ,
  • ZOU Yuxiao ,
  • LI Erna
Expand
  • 1(College of Food Science, South China Agricultural University, Guangzhou 510642, China)
    2(Sericultural & Agri-Food Research Institute, Guangdong Academy of Agricultural Sciences, Key Laboratory of Functional Foods, Ministry of Agriculture and Rural Affairs, Guangdong Key Laboratory of Agricultural Products Processing, Guangzhou 510610, China)

Received date: 2025-02-04

  Revised date: 2025-06-01

  Online published: 2026-03-13

摘要

为了研究桑椹低聚半乳糖(mulberry galacto-oligosaccharide,MGO)对鼠李糖乳酪杆菌EN-08胞内蛋白质表达的影响,采用串联质量标签(tandem mass tags,TMT)定量蛋白质组学技术进行分析。将鼠李糖乳酪杆菌EN-08分别接种到含MGO、桑椹多糖(mulberry polysaccharide,MPS)和低聚半乳糖(galactooligosaccharides,GOS)的MRS培养基中发酵培养。培养18 h后提取所有样品的蛋白质进行TMT技术鉴定,采用相关性热图和主成分分析观察各组相关性,发现添加MGO和MPS后,鼠李糖乳酪杆菌EN-08全蛋白组成发生变化。在MGO组中鉴定出192个差异蛋白,其中上调蛋白有110个,下调蛋白有82个。KEGG功能注释显示,上调差异蛋白的功能主要集中在碳水化合物代谢和膜运输方面,分别有17个和16个上调蛋白参与。KEGG富集分析显示,上调差异蛋白质显著富集于果糖和甘露糖代谢以及磷酸转移酶系统通路,富集因子分别为0.219 5和0.228 6。分析结果表明,MGO主要从碳水化合物代谢以及环境信息处理这两方面影响菌株的生命活动。说明MGO可作为有潜力的碳源,促进菌体代谢调控,为鼠李糖乳酪杆菌EN-08的高密度工业化生产提供参考。

本文引用格式

陈心如 , 李倩 , 庞道睿 , 邢东旭 , 邹宇晓 , 黎尔纳 . 基于蛋白质组学解析桑椹低聚半乳糖通过碳水化合物代谢调控鼠李糖乳酪杆菌EN-08增殖的机制[J]. 食品与发酵工业, 2026 , 52(3) : 121 -128 . DOI: 10.13995/j.cnki.11-1802/ts.042496

Abstract

To investigate the effect of mulberry galacto-oligosaccharide (MGO) on the intracellular protein expression of Lacticaseibacillus rhamnosus EN-08, Tandem Mass Tags (TMT) quantitative proteomics technology was employed for analysis.L. rhamnosus EN-08 was inoculated into MRS media containing mulberry galacto-oligosaccharide (MGO), mulberry polysaccharide (MPS), and galactooligosaccharides (GOS) for fermentation, respectively.After 18 hours of cultivation, proteins were extracted from all samples for TMT-based identification.Correlation heatmaps and principal component analysis revealed changes in the overall protein composition of L. rhamnosus EN-08 with the addition of MGO and MPS.A total of 192 differentially expressed proteins were identified in the MGO group, including 110 up-regulated proteins and 82 down-regulated proteins.KEGG functional annotation revealed that the up-regulated differential proteins primarily involved carbohydrate metabolism and membrane transport, with 17 and 16 upregulated proteins, respectively.KEGG enrichment analysis showed that the up-regulated differential proteins were significantly enriched in fructose and mannose metabolism and phosphotransferase system pathways, with rich factors of 0.219 5 and 0.228 6, respectively.The analysis results suggested that MGO primarily influenced the metabolic activities of the strain through carbohydrate metabolism and environmental information processing.This indicated that MGO had the potential to serve as a carbon source, promoting bacterial metabolic regulation and providing a reference for the high-density industrial production of L. rhamnosus EN-08.

参考文献

[1] 单强, 王雪, 朱要宏, 等.鼠李糖乳杆菌抗炎机制及其在防治家畜疾病中的应用前景[J].畜牧兽医学报, 2023, 54(11):4537-4550.
SHAN Q, WANG X, ZHU Y H, et al.Application prospect of anti-inflammatory mechanism of Lactobacillus rhamnosus and its prevention and treatment in livestock diseases[J].Acta Veterinaria et Zootechnica Sinica, 2023, 54(11):4537-4550.
[2] SANKLECHA M, VERMA L, PAI U, et al.Lactobacillus rhamnosus GG evaluation in acute diarrhea (LEAD):An observational study[J].Cureus, 2022, 14(4):e24594.
[3] 刘波, 刘通通, 张祥奎, 等.益生元及其应用研究进展[J].食品与药品, 2021, 23(5):485-492.
LIU B, LIU T T, ZHANG X K, et al.Research progress of prebiotics and their applications[J].Food and Drug, 2021, 23(5):485-492.
[4] 杨诗沅, 邹宇晓, 胡腾根, 等.桑枝低聚糖促乳酸菌生长活性优化研究[J].食品科学技术学报, 2020, 38(2):66-72.
YANG S Y, ZOU Y X, HU T G, et al.Optimization of growth promoting activity of Ramulus mori oligosaccharide on lactic acid bacteria[J].Journal of Food Science and Technology, 2020, 38(2):66-72.
[5] 李艳丽, 许少春, 柳永, 等.低聚木糖的制备及其对益生菌体外增殖的作用[J].浙江大学学报(农业与生命科学版), 2011, 37(3):245-251.
LI Y L, XU S C, LIU Y, et al.Preparation of xylo-oligosaccharides and its effect on in vitro proliferation of probiotics[J].Journal of Zhejiang University (Agriculture and Life Sciences), 2011, 37(3):245-251.
[6] MA X Q, ZHENG Z C, WANG Q, et al.The modulation effect of lotus (Nelumbo nucifera Gaertn.) seeds oligosaccharides with different structures on intestinal flora and action mode of growth effects on Bifidobacterium in vivo and in vitro[J].Food Chemistry, 2023, 419:136057.
[7] CHENG Y X, ZHAO R Q, QIAO M W, et al.The pea oligosaccharides could stimulate the in vitro proliferation of beneficial bacteria and enhance anti-inflammatory effects via the NF-κB pathway[J].Foods, 2024, 13(4):626.
[8] LI E N, YANG S Y, ZOU Y X, et al.Purification, characterization, prebiotic preparations and antioxidant activity of oligosaccharides from mulberries[J].Molecules, 2019, 24(12):2329.
[9] ASLAM B, BASIT M, NISAR M A, et al.Proteomics:Technologies and their applications[J].Journal of Chromatographic Science, 2017, 55(2):182-196.
[10] JIANG Y Y, ZHANG M, ZHANG Y, et al.Calcium (Ca(2+))-regulated exopolysaccharide biosynthesis in probiotic Lactobacillus plantarum K25 as analyzed by an omics approach[J].Journal of Dairy Science, 2021, 104(3):2693-2708.
[11] VEGA-SAGARDÍA M, CABEZÓN E C, DELGADO J, et al.Screening microbial interactions during inulin utilization reveals strong competition and proteomic changes in Lacticaseibacillus paracasei M38[J].Probiotics and Antimicrobial Proteins, 2024, 16(3):993-1011.
[12] VEGA-SAGARDÍA M, DELGADO J, RUIZ-MOYANO S, et al.Proteomic analyses of Bacteroides ovatus and Bifidobacterium longum in xylan bidirectional culture shows sugar cross-feeding interactions[J].Food Research International, 2023, 170:113025.
[13] LI E N, ZHU Q L, PANG D R, et al.Analysis of Lactobacillus rhamnosus GG in mulberry galacto-oligosaccharide medium by comparative transcriptomics and metabolomics[J].Frontiers in Nutrition, 2022, 9:853271.
[14] JIANG P F, CHEN X W, QIAN L, et al.Integrated transcriptomic and proteomic analysis of the physiological changes of the liver in domesticated Eurasian perch, Perca fluviatilis[J].Comparative Biochemistry and Physiology Part D:Genomics and Proteomics, 2022, 41:100957.
[15] 许家齐, 郑新飞, 冯燕, 等.基于蛋白质组学研究冷冻干燥对副干酪乳杆菌PC-01细胞活性的影响[J].中国食品学报, 2023, 23(3):177-186.
XU J Q, ZHENG X F, FENG Y, et al.Effects of freeze-drying on the activity of Lactobacillus paracasei PC-01 cells based on proteomics[J].Journal of Chinese Institute of Food Science and Technology, 2023, 23(3):177-186.
[16] 冯佳, 吴昊, 王斌斌, 等.全局调控因子CodY在益生菌中的研究进展[J].食品与发酵工业, 2016, 42(10):227-233.
FENG J, WU H, WANG B B, et al.Advances in research on global regulatory factor CodY of probiotics[J].Food and Fermentation Industries, 2016, 42(10):227-233.
[17] WARDMAN J F, BAINS R K, RAHFELD P, et al.Carbohydrate-active enzymes (CAZymes) in the gut microbiome[J].Nature Reviews Microbiology, 2022, 20(9):542-556.
[18] CUI Y H, WANG M H, ZHENG Y K, et al.The carbohydrate metabolism of lactiplantibacillus plantarum[J].International Journal of Molecular Sciences, 2021, 22(24):13452.
[19] VAN VEEN H W, KONINGS W N.The ABC family of multidrug transporters in microorganisms[J].Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1998, 1365(1-2):31-36.
[20] QIU Y J, QIU Z Y, XIA J, et al.Co-expression of xylose transporter and fructose-bisphosphate aldolase enhances the utilization of xylose by Lactococcus lactis IO-1[J].Applied Biochemistry and Biotechnology, 2023, 195(2):816-831.
[21] SCHOENENBERGER B, KIND S, MEIER R, et al.Efficient biocatalytic synthesis of D-tagatose 1, 6-diphosphate by LacC-catalysed phosphorylation of D-tagatose 6-phosphate[J].Biocatalysis and Biotransformation, 2020, 38(1):53-63.
[22] SINGH S, GOLA C, SINGH B, et al.D-galactonate metabolism in enteric bacteria:A molecular and physiological perspective[J].Current Opinion in Microbiology, 2024, 81:102524.
[23] ZHOU Y C, BU Z Y, QIAN J, et al.The UTP-glucose-1-phosphate uridylyltransferase of Brucella melitensis inhibits the activation of NF-κB via regulating the bacterial type IV secretion system[J].International Journal of Biological Macromolecules, 2020, 164:3098-3104.
[24] NOBELMANN B, LENGELER J W.Molecular analysis of the gat genes from Escherichia coli and of their roles in galactitol transport and metabolism[J].Journal of Bacteriology, 1996, 178(23):6790-6795.
文章导航

/

〈 〉